Food DNA isolation miniprep kit: Manual

5. Column washing 1
Wash the column with 500µl Wash Buffer 1 and centrifuge at 5,000 x g for 1 min.
Discard through.
Ensure that ethanol has been added into the Wash Buffer 1 before use (refer to Reconstitution of
Solutions).
6. Column washing 2
Wash the column with 500µl Wash Buffer 2 and centrifuge at 5,000 x g for 1 min.
Discard Flow through. Wash the column again with 500µl Wash Buffer 2 and centrifuge
at maximum speed for 3 min.
Ensure that ethanol has been added into the Wash Buffer 2 before use (refer to Reconstitution of
Solutions). Ensure centrifugation for 3 min to remove ethanol completely.
7. DNA Elution
Place the column into a clean microcentrifuge tube. Add 50-100µl of preheated Elution
Buffer, TE buffer or sterile water directly onto column membrane and stand for 2 min.
Centrifuge at 5,000 x g for 1 min to elute DNA. Store DNA at 4°C or -20°C.
Ensure that the Elution Buffer is dispensed directly onto the center of the membrane for complete
elution. TEBuffer can also elute DNA although EDTA may inhibit subsequent enzymatic
reactions. If water is used for eluting DNA, maximum elution efficiency is achieved between
pH7.0 and 8.5. Store DNA at -20°C as DNA may degrade in the absence of a buffering agent.
Note: The recommended amplicon length for PCR analysis is